Methods of using cyclooxygenase-prostacyclin synthase fusion gene

Abstract

An effective amount of a composition comprising (i) a plasmid having a cyclooxygenase-prostacyclin synthase fusion gene, and (ii) a carrier fluid for use in treating an individual having a vascular disease or at risk of developing a vascular disease. A composition comprising a carrier fluid; and a DNA sequence encoding for a triple catalytic enzyme, a cDNA sequence encoding for a triple catalytic enzyme, a plasmid comprising a DNA sequence encoding for a triple catalytic enzyme, a fusion gene encoding for a triple catalytic enzyme, a cyclooxygenase-prostacyclin synthase fusion gene, or combinations thereof, for use in treating an individual having a vascular disease or at risk of developing a vascular disease.

1 . An effective amount of a composition comprising (i) a plasmid having a cyclooxygenase-prostacyclin synthase fusion gene, and (ii) a carrier fluid for use in treating an individual having a vascular disease or at risk of developing a vascular disease. 
     
     
         2 . The composition of  claim 1  wherein the vascular disease comprises: pulmonary arterial hypertension, peripheral arterial disease, peripheral vascular disease, chronic obstructive pulmonary disease, ischemia, limb ischemia, critical limb ischemia, Reynaud's syndrome, ischemic stroke, myocardial infarction, systemic hypertension, stroke, subarachnoid hemorrhage, or combinations thereof. 
     
     
         3 . The composition of  claim 1  wherein the plasmid comprises a DNA sequence encoding for a triple catalytic enzyme. 
     
     
         4 . The composition of  claim 3  wherein the triple catalytic enzyme is characterized by a formula COX-linker-PGIS, wherein COX comprises a cyclooxygenase (COX) amino acid sequence; PGIS is prostacyclin synthase; and the linker comprises from about 10 to about 22 amino acid residues of a transmembrane sequence; wherein the linker is disposed between the COX and PGIS, and wherein the linker directly connects the COX to the PGIS. 
     
     
         5 . The composition of  claim 4  wherein the triple catalytic enzyme is characterized by a formula COX-1-10aa-PGIS; wherein COX-1 is cyclooxygenase isoform-1; the linker comprises a 10 amino acid (10aa) transmembrane sequence; and PGIS is prostacyclin synthase. 
     
     
         6 . The composition of  claim 1  wherein the plasmid comprises a constitutively active mammalian promoter. 
     
     
         7 . The composition of  claim 1  wherein the plasmid comprises an inducible promoter. 
     
     
         8 . The composition of  claim 1  wherein the plasmid comprises a human muscle specific promoter, human muscle creatinine kinase promoter, human a-skeletal actin promoter, human desmin promoter, human troponin I promoter, human cytomegalovirus promoter, antibiotic-inducible promoter, tetracycline-inducible promoter, doxycycline-inducible promoter, bacitracin-inducible promoter, vancomycin-inducible promoter, and isopropyl beta-D-thiogalactopyranoside (IPTG)-inducible promoter. 
     
     
         9 . The composition of  claim 1  administered via an intramuscular injection. 
     
     
         10 . The composition of  claim 1  wherein the composition is injected into an ischemic tissue. 
     
     
         11 . The composition of  claim 1  wherein the vascular disease comprises pulmonary arterial hypertension, and wherein the composition is injected into a brachial skeletal muscle. 
     
     
         12 . The composition of  claim 1  wherein the plasmid enters a host cell upon administering said composition to said individual, and wherein said host cell overexpresses prostacyclin (PGI2). 
     
     
         13 . The composition of  claim 1  wherein the carrier fluid comprises phosphate buffer saline. 
     
     
         14 . The composition of  claim 1  wherein the plasmid is present within the composition in an amount of from about 0.4 mg/ml to about 500 mg/ml. 
     
     
         15 . The composition of  claim 1  excluding a viral vector. 
     
     
         16 . A composition comprising a carrier fluid; and a DNA sequence encoding for a triple catalytic enzyme, a cDNA sequence encoding for a triple catalytic enzyme, a plasmid comprising a DNA sequence encoding for a triple catalytic enzyme, a fusion gene encoding for a triple catalytic enzyme, a cyclooxygenase-prostacyclin synthase fusion gene, or combinations thereof, for use in treating an individual having a vascular disease or at risk of developing a vascular disease. 
     
     
         17 . The composition of  claim 16  administered via an intramuscular injection into an ischemic tissue. 
     
     
         18 . The composition of  claim 16  wherein the triple catalytic enzyme is characterized by a formula COX-1-10aa-PGIS; wherein COX-1 is cyclooxygenase isoform-1; the linker comprises a 10 amino acid (10aa) transmembrane sequence; and PGIS is prostacyclin synthase. 
     
     
         19 . A composition comprising a plasmid comprising a cyclooxygenase-prostacyclin synthase fusion gene, wherein the plasmid comprises a human muscle specific promoter, and phosphate buffer saline, for use in treating an individual having pulmonary arterial hypertension or at risk of developing pulmonary arterial hypertension, wherein the plasmid is present within the composition in an amount of from about 0.4 mg/ml to about 500 mg/ml; and wherein the composition is administered via an intramuscular injection into a brachial skeletal muscle. 
     
     
         20 . A composition for prostacyclin (PGI2) delivery, wherein the composition for PGI2 delivery comprises: a carrier fluid; and a DNA sequence encoding for a triple catalytic enzyme, a cDNA sequence encoding for a triple catalytic enzyme, a plasmid comprising a DNA sequence encoding for a triple catalytic enzyme, a fusion gene encoding for a triple catalytic enzyme, a cyclooxygenase-prostacyclin synthase fusion gene, or combinations thereof. 
     
     
         21 . The composition of  claim 20 , wherein the plasmid is present within the composition in an amount of from about 0.4 mg/ml to about 500 mg/ml.